Ďurč Pavol, Foret František, Pokojová Eva, Homola Lukáš, Skřičková Jana, Herout Vladimír, Dastych Milan, Vinohradská Hana, Kubáň Petr
Department of Bioanalytical Instrumentation, CEITEC Masaryk University, Veveří 97, 602 00, Brno, Czech Republic.
Department of Chemistry, Masaryk University, Kotlářská 2, 611 37, Brno, Czech Republic.
Anal Bioanal Chem. 2017 May;409(14):3507-3514. doi: 10.1007/s00216-017-0318-6. Epub 2017 Mar 29.
A new approach for sweat analysis used in cystic fibrosis (CF) diagnosis is proposed. It consists of a noninvasive skin-wipe sampling followed by analysis of target ions using capillary electrophoresis with contactless conductometric detection (C4D). The skin-wipe sampling consists of wiping a defined skin area with precleaned cotton swab moistened with 100 μL deionized water. The skin-wipe sample is then extracted for 3 min into 400 μL deionized water, and the extract is analyzed directly. The developed sampling method is cheap, simple, fast, and painless, and can replace the conventional pilocarpine-induced sweat chloride test commonly applied in CF diagnosis. The aqueous extract of the skin-wipe sample content is analyzed simultaneously by capillary electrophoresis with contactless conductometric detection using a double opposite end injection. A 20 mmol/L L-histidine/2-(N-morpholino)ethanesulfonic acid and 2 mmol/L 18-crown-6 at pH 6 electrolyte can separate all the major ions in less than 7 min. Skin-wipe sample extracts from 30 study participants-ten adult patients with CF (25-50 years old), ten pediatric patients with CF (1-15 years old), and ten healthy control individuals (1-18 years old)-were obtained and analyzed. From the analyzed ions in all samples, a significant difference between chloride and potassium concentrations was found in the CF patients and healthy controls. We propose the use of the Cl/K ratio rather than the absolute Cl concentration and a cutoff value of 4 in skin-wipe sample extracts as an alternative to the conventional sweat chloride analysis. The proposed Cl/K ion ratio proved to be a more reliable indicator, is independent of the patient's age, and allows better differentiation between non-CF individuals and CF patients having intermediate values on the Cl sweat test. Figure New approach for cystic fibrosis diagnosis based on skin-wipe sampling of forearm and analysis of ionic content (Cl/K ratio) in skin-wipe extracts by capillary electrophoresis with contactless conductometric detection.
提出了一种用于囊性纤维化(CF)诊断的汗液分析新方法。该方法包括无创皮肤擦拭采样,随后使用毛细管电泳结合非接触式电导检测(C4D)对目标离子进行分析。皮肤擦拭采样是用蘸有100μL去离子水的预清洁棉签擦拭特定的皮肤区域。然后将皮肤擦拭样本在400μL去离子水中提取3分钟,提取物直接进行分析。所开发的采样方法廉价、简单、快速且无痛,可替代CF诊断中常用的传统毛果芸香碱诱导汗液氯化物测试。使用双端反向进样,通过毛细管电泳结合非接触式电导检测同时分析皮肤擦拭样本提取物中的水相成分。在pH值为6的20 mmol/L L-组氨酸/2-(N-吗啉代)乙磺酸和2 mmol/L 18-冠-6电解质中,所有主要离子可在不到7分钟内分离。获取并分析了30名研究参与者的皮肤擦拭样本提取物,其中包括10名成年CF患者(25至50岁)、10名儿科CF患者(1至15岁)和10名健康对照个体(1至18岁)。在所有样本分析的离子中,CF患者和健康对照者的氯化物和钾浓度存在显著差异。我们建议使用皮肤擦拭样本提取物中的Cl/K比值而非绝对Cl浓度,并将截止值设为4,作为传统汗液氯化物分析的替代方法。所提出的Cl/K离子比值被证明是一个更可靠的指标,与患者年龄无关,并且能够更好地区分非CF个体和汗液氯化物测试值处于中间范围的CF患者。图 基于前臂皮肤擦拭采样及通过毛细管电泳结合非接触式电导检测分析皮肤擦拭提取物中离子含量(Cl/K比值)的囊性纤维化诊断新方法。